Improvement in the fermentation quality of Italian ryegrass silage by ensiling with combined lactic acid bacteria inoculants at low temperature

doi:10.11686/cyxb2024226

Abstract

Abstract:

Temperature is an important factor affecting ensilage. In cold regions， low temperature can be an adverse environmental condition for ensiling. However， few studies have focused on improving the ensiling process at low temperature to enhance silage quality. The aims of this study were： 1） To characterize the lactic acid bacteria （LAB） strains Pediococcus acidilactici LOG9， Pediococcus pentosaceus LO7， Lentilactobacillus coryniformis subsp. LM8， and Lactiplantibacillus plantarum M1， which were isolated from the Tibetan Plateau. 2） Determine the effect of adding combinations of these strains on the quality of Italian ryegrass （Lolium multiflorum） silage produced at three temperatures （10， 15， 25 ℃）. The isolated strains were subjected to morphological， physiological， and biochemical analyses. Four combined inoculants （LO7+LM8， LO7+M1， LOG9+LM8， and LOG9+M1） were added to Italian ryegrass （at 1×10⁵ cfu·g^-1 fresh weight）. These mixtures were then ensiled for 60 days in laboratory silos （1 L） at different ambient temperatures （10， 15， 25 ℃）. All the isolates were able to grow normally at 5-20 ℃， pH 3.5-7.0， with NaCl concentrations of 3.0% and 6.5% w/w. Compared with the three corresponding controls， all the combined LAB inoculants improved the quality of Italian ryegrass silage， as indicated by markedly higher （P<0.05） lactic acid （LA） contents， higher lactic acid/acetic acid （LA/AA）， lower pH and ammonia nitrogen （NH₃-N） contents， and lower counts of undesirable microorganisms. With ensilage at 10 and 15 ℃， LOG9+LM8/M1 inoculants performed better than the LO7+LM8/M1 inoculants， as demonstrated by the distinctly higher （P<0.05） LA contents and LA/AA， and lower NH₃-N contents. Compared with the silage produced with LOG9+M1， that produced with LOG9+LM8 had significantly （P<0.05） higher LA contents and LAB counts， and lower NH₃-N contents. Therefore， the combined inoculant LOG9+LM8 is recommended as the starter culture for producing Italian ryegrass silage at low temperature.

Key words: fermentation quality, combined lactic acid bacteria inoculant, Italian ryegrass, silage, low temperature

Si-ran WANG, Bei-yi LIU, Ji-peng TIAN, Yun-hui CHENG, Neng-xiang XU, Wen-jie ZHANG, Xin WANG, Cheng-long DING. Improvement in the fermentation quality of Italian ryegrass silage by ensiling with combined lactic acid bacteria inoculants at low temperature[J]. Acta Prataculturae Sinica, 2025, 34(5): 159-170.

Figures/Tables 9

References 37

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项目Items	LOG9	LO7	LM8	M1
形状Shape	球菌Cocci	球菌Cocci	杆菌Rod	杆菌Rod
发酵类型Fermentation type	同型Homo	同型Homo	同型Homo	同型Homo
革兰氏染色Gram stain	+	+	+	+
过氧化氢酶活性Catalase activity	-	-	-	-
葡萄糖产气试验Gas from glucose	-	-	-	-
温度Temperature （℃）
5	+	+	+	+
10	+	+	+	+
15	+	+	+	+
20	+	+	+	+
pH
3.0	W	-	-	W
3.5	+	+	+	+
4.0	+	+	+	+
4.5	+	+	+	+
5.0	+	+	+	+
6.0	+	+	+	+
7.0	+	+	+	+
盐度NaCl （%）
3.0	+	+	+	+
6.5	+	+	+	+

项目Items	LOG9	LO7	LM8	M1
形状Shape	球菌Cocci	球菌Cocci	杆菌Rod	杆菌Rod
发酵类型Fermentation type	同型Homo	同型Homo	同型Homo	同型Homo
革兰氏染色Gram stain	+	+	+	+
过氧化氢酶活性Catalase activity	-	-	-	-
葡萄糖产气试验Gas from glucose	-	-	-	-
温度Temperature （℃）
5	+	+	+	+
10	+	+	+	+
15	+	+	+	+
20	+	+	+	+
pH
3.0	W	-	-	W
3.5	+	+	+	+
4.0	+	+	+	+
4.5	+	+	+	+
5.0	+	+	+	+
6.0	+	+	+	+
7.0	+	+	+	+
盐度NaCl （%）
3.0	+	+	+	+
6.5	+	+	+	+

项目Items	LOG9	LO7	LM8	M1	项目Items	LOG9	LO7	LM8	M1
L-阿拉伯糖L-arabinose	++	++	-	++	蜜二糖Melibiose	++	++	++	++
核糖Ribose	++	++	-	++	蔗糖Saccharose	++	++	-	++
鼠李糖Rhamnose	+	-	-	-	海藻糖Trehalose	++	++	-	++
甘露醇Mannitol	++	-	++	++	松三糖Melezitose	++	-	-	++
山梨醇Sorbitol	++	-	++	++	D-棉籽糖D-raffinose	++	-	++	++
α-甲基-D-甘露糖苷 α-methyl-D-mannoside	++	-	-	++	β-龙胆二糖β-gentiobiose	+	+	-	+
α-甲基-D-甘露糖苷 α-methyl-D-mannoside	++	-	-	++	D-松二糖D-turanose	++	-	-	++
苦杏仁苷Amygdaline	++	++	-	++	L-岩藻糖L-fucose	-	-	+	-
七叶灵Esculine	++	++	-	++	D-阿拉伯糖醇D-arabitol	W	-	-	W
纤维二糖Cellobiose	++	++	-	++	葡萄糖酸盐Gluconate	W	W	-	+
乳糖Lactose	++	++	-	++

项目Items	LOG9	LO7	LM8	M1	项目Items	LOG9	LO7	LM8	M1
L-阿拉伯糖L-arabinose	++	++	-	++	蜜二糖Melibiose	++	++	++	++
核糖Ribose	++	++	-	++	蔗糖Saccharose	++	++	-	++
鼠李糖Rhamnose	+	-	-	-	海藻糖Trehalose	++	++	-	++
甘露醇Mannitol	++	-	++	++	松三糖Melezitose	++	-	-	++
山梨醇Sorbitol	++	-	++	++	D-棉籽糖D-raffinose	++	-	++	++
α-甲基-D-甘露糖苷 α-methyl-D-mannoside	++	-	-	++	β-龙胆二糖β-gentiobiose	+	+	-	+
α-甲基-D-甘露糖苷 α-methyl-D-mannoside	++	-	-	++	D-松二糖D-turanose	++	-	-	++
苦杏仁苷Amygdaline	++	++	-	++	L-岩藻糖L-fucose	-	-	+	-
七叶灵Esculine	++	++	-	++	D-阿拉伯糖醇D-arabitol	W	-	-	W
纤维二糖Cellobiose	++	++	-	++	葡萄糖酸盐Gluconate	W	W	-	+
乳糖Lactose	++	++	-	++

菌株 Strain	登录号 Accession number	16S rRNA基因测序数据（近源种） 16S rRNA gene sequencing data （closest relative）	相似度 Similarity （%）
LOG9	KJ779095	乳酸片球菌DSM 20284 P. acidilactici DSM 20284	99
LO7	KJ779092	戊糖片球菌DSM 20336 P. pentosaceus DSM 20336	99
LM8	KJ779090	棒状乳杆菌亚种torquens 30 L. coryniformis subsp. torquens 30	99
M1	KJ779098	植物乳杆菌JCM 1149 L. plantarum JCM 1149	99